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Nucleic Acids Research, 1988, Vol. 16, No. 20 9457-9475
© 1988

Articles

Splicing and spliceosome formation of the yeast MATa1 transcript require a minimum distance from the 5' splice site to the internal branch acceptor site

Karl Köhrer and Horst Domdey*

Laboratorium für Molekulare Biologie, Genzentrum der Ludwig-Maximilians, Universität München D-8033 Martinsried, FRG

*To whom correspondence should be addressed

Received August 8, 1988. Accepted September 23, 1988.

Small deletions of 6, 7, and 12 nucleotides introduced between the 5' splice site and the internal branch acceptor site of the first intron of the yeast MATaI gene completely abolish accurate splicing in vitro in these constructs. Splicing only occurs at an alternative 5' splice site which was found in the first exon of the MATaI gene and which is used both in vivo and in vitro. The splicing defect cannot be cured by expanding the distance from the branch point to the 3' splice site. If the alternative 5' splice site is deleted as well in these constructs, neither spliced products nor spliceosomes are formed. Our findings especially lead to the conclusion that a minimum distance between the 5' splice site and the internal branch acceptor site of the intron is required for the formation of splicing complexes and for accurate splicing.


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